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A common SLC26A6 haplotype is not associated with chronic pancreatitis
S78
Abstracts / Pancreatology 14 (2014) S1eS129
Background: There are no extensive data on the comparative evaluation of 25-hydroxyvitamin D (VIT D) serum levels in patients with chronic benign and malignant pancreatic diseases. Aims: To evaluate serum VIT D levels in patients with chronic pancreatitis (CP), pancreatic adenocarcinoma (PA) and in healthy subjects (HS). Patients & methods: Sixty-nine consecutive subjects were studied: 20 CP patients (12 M, 8 F; mean age 56.0 yrs, range 25-74; 14 were alcohol drinkers and 12, smokers; 12 had calcifications and 5 diabetes mellitus), 20 PA (11 M, 9 F; age 63.5 yrs, range 47-80; 5 patients were alcohol drinkers, 3 were smokers; 11 had diabetes); 29 HS (13 M, 16 F; mean age 59.5 yrs, range 44-75). Serum VIT D levels were measured using chemiluminescence assay . VIT D levels were normally distributed and ANOVA test was applied. Fecal pancreatic elastase-1 (FE) was also measured in patients. Results: The 3 groups were comparable for sex (P¼0.554) and age (P¼0.107); 14 CP patients and 1 PA had exocrine pancreatic insufficiency (P<0.001). Serum VIT D was significantly different among the 3 groups of patients studied (mean±SD; CP: 10.9±8.1 ng/mL, PA: 12.5±5.6 ng/mL, HS: 23.6±8.3 ng/mL; P<0.001). CP and PA patients had similar VIT D serum levels (P¼0.258) and they were significantly lower than in HS (P<0.001). At univariate analysis only the type of the disease (i.e. CP or PA) was significantly associated with low VIT D levels (P¼0.036). Conclusion: Low VIT D levels are associated with chronic diseases of the pancreas, but the mechanism should be better elucidated.
Aims: This study aims to distinguish nutritional status of TP-IAT patients pre- and post-surgery. Patients & methods: Nutrition parameters (BMI, vitamin D3, vitamin A, and vitamin E levels) were obtained from an IRB-approved prospective database collected for TP-IAT patients at a single institution. Two-tailed paired T-test and two-tailed student T-test for unequal variances were used for data analysis. Results: Weight loss was observed from pre-op to 6 months post-op (n¼111) from mean BMI (26.1kg/m2+/-6.6 to 23.0kg/m2+/-4.9, p<0.0001). There was little change in weight from 6 months to 1 year post-op. BMI at 2 and 3 years post-op were also unchanged. Average vitamin D3 levels dropped significantly pre-op to 6 months post-op (27.8ng/ml+/-10.5 to 24.4ng/ml+/-10.2, p¼0.02). Levels also dropped from 6 months post-op to 1 year post-op (24.4ng/ml+/-10.2 to 22.5ng/ml+/-11.8) indicating moderate deficiency after TP-IAT. Despite having fewer data at 2 and 3 years post-op, mean vitamin D3 levels indicated prolonged vitamin D3 deficiency (20.3ng/ml+/-11.8, 21.6ng/ml+/-15.8). Mean levels of a subpopulation (n¼55) for vitamin A and vitamin E were normal (0.34mg/L+/-0.15 and 7.4mg/L+/-3.1) at time points ranging from 6 months post-op to 3 years post-op. However, vitamin A and vitamin E deficiencies were observed in 45% and 29% of subjects respectively. Conclusion: Nutritional compromise is a notable problem in patients after TP-IAT. Although nutritional risk may improve over time, these patients require long-term follow-up.
T-098. Clinical significance of sarcopenia as a predictor of prognosis in patients who underwent pancreaticoduodenectomy. Fumiaki Watanabe, Hiroshi Noda, Toshiki Rikiyama Jichi Medical University Saitama Medical Center, Surgery, Japan Background: Sarcopenia was identified recently as a poor prognostic factor in patients with cancer. The present study investigated the influence of sarcopenia on the perioperative clinical outcomes and long-term survival of the patients who underwent pancreaticoduodenectomy(PD). Aims: Patients were assigned to one of two groups according to the presence or absence of sarcopenia. Clinicopathological, surgical outcome and long-term survival data were analysed. Patients & methods: Data were collected retrospectively for 123 consecutive patients who underwent PD for pancreatic head cancer, carcinoma of vater, common bile duct cancer, and IPMN with a curative intent between April 2008 and December 2012. Patients were divided into two groups according to the presence or absence of sarcopenia, assessed by computed tomographic measurement of muscle mass at the level of the third lumbar vertebra (N. Harimoto, et al. British Journal of Surgery 2013; 100: 1523e30). Clinicopathologica characteristics, postoperative outcomes and long-term survival data were compared between two groups. Results: Sarcopenia was present in 60 (48.8 per cent) of 123 patients, and was significantly correlated with female sex. In patients with and without sarcopenia, the rate of morbidities was 30.0 and 36.5 per cent, the duration postoperative hospital stay was 29.0 and 29.3 days, and the 5-year overall survival rate was 54.3 and 76.7 per cent, respectively (p¼0.067). Conclusion: It might be possible that sarcopenia was a predictive factor for poor prognosis of patients who underwent PD.
T-099. Auto-islet cell transplantation: What is happening to weight and fatsoluble vitamin status? Maria Nestleroad, Kelley Martin, Elizabeth Shufford, Stefanie Owczarski, David Adams, Katherine Morgan Medical University of South Carolina, Department of Gastrointestinal and Laparoscopic Surgery, United States Background: Limited data links chronic pancreatitis to nutritional compromise after Total pancreatectomy and islet auto-transplantation (TPIAT).
T-100. A common SLC26A6 haplotype is not associated with chronic pancreatitis zs a, Eszter Hegyi a, Bala zs Csaba N n Hritz c, Judit Anita Bala emeth b, Istva c c d d € Gervain , Ferenc Izbeki , Attila Szepes , Gyorgy Gyimesi , Dezs g, Zsolt Vincze e, Judit Bajor f, Adrienn Csiszko Kelemen e, Aron s Bod h, Andrea Pa rniczky i, La sztity Nata lia i, Szentkereszty g, Barnaba bor Veres j, Richa rd Szmola k, Akos Pap k , Judit Czelecz l, Csilla Andorka j, Ga nos Nova k n, Gyula Farkas o, La szlo s nos Sümegi m, Ja Czako a, Tama Ja a a a th b, P eter Takacs , Zoltan Rakonczay , Jozsef Maleth , Miklos Sahin-To Hegyi a a
First Department of Medicine, University of Szeged, Szeged, Hungary Department of Molecular and Cell Biology, Boston University, Boston, United States c r Megyei Szent Gyo € rgy Hospital, Sze kesfehe rva r, Hungary Feje d t, Hungary Bacs-Kiskun County Municipality Hospital, Kecskeme e cs, Hungary Department of Surgery, University of Pe f cs, Hungary Department of Internal Medicine, University of Pe g Department of Surgery, University of Debrecen, Hungary h n Hospital, Szentes, Hungary Dr. Bugyi Istva i Heim P al Children?s Hospital, Budapest, Hungary j Semmelweis University,Paediatric Department, Budapest, Hungary k National Institute of Oncology, Budapest, Hungary, Hungary l Bethasda Children?s Hospital, Budapest, Hungary m B-A-Z County Hopspital, Miskolc, Hungary n ndy Ka lma n County Hospital, Gyula, Hungary 9Pa o Department of Surgery, University of Szeged, Hungary b
Background: Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) mutations are established risk factors for chronic pancreatitis (CP). CFTR variants increase disease risk by causing impairment of pancreatic ductal bicarbonate secretion. However, the role of genetic variations in the bicarbonate secreting SLC26A6 anion transporter has remained largely unexplored so far. Aims: Our aim was to investigate the role of the SLC26A6 gene in CP. Patients & methods: 96 subjects with CP (cases) and 99 subjects with no pancreatic disease (controls) were recruited from the Hungarian National Pancreas Registry. In a discovery cohort of 30 idiopathic CP cases the entire SLC26A6 coding sequence, including 21 exons and the exonintron boundaries were amplified and sequenced. Further genotyping of p.V206M and p.P397P mutations in CP and controls was carried out by RFLP.
Abstracts / Pancreatology 14 (2014) S1eS129
Results: Sequencing analysis of the discovery cohort revealed four common mutations: intronic mutations c.23+71_23+103del, c.183-4C>A and c.1134+32C>A; and exonic missense mutation p.V206M. . These four mutations were found in linkage disequilibrium indicating a conserved haplotype. We found this haplotype in 18 heterozygous and 2 homozygous cases, and in 24 heterozygous and 2 homozygous controls (allele frequency 11.4% and 14.1% respectively). A synonymous mutation p.P397P was also detected in a single case. Conclusion: We found a novel, common haplotype in the SLC26A6 gene, which did not show association with CP. Supported by TAMOP and OTKA.
T-101. Expression of aquaporins in pancreatic ductal cells n Rakonczay, Jr. b, Viktoria Venglovecz a, Lajos V. Kem eny b, Zolta szlo s c, P Puska eter Hegyi b Agnes Zvara c, La a University of Szeged, Department of Pharmacology and Pharmacotherapy, Hungary b University of Szeged, First Department of Medicine, Hungary c Biological Research Centre, Laboratory of Functional Genomics, Hungary
Background: Acute pancreatitis (AP) is a multicellular disease in which pancreatic ductal cells play an important role. Toxic agents inducing AP inhibit pancreatic ductal bicarbonate secretion, however, no information is available concerning their effects on the regulation of aquaporins (AQPs). Aims: The aim of this study was to investigate the effects of bile acids, ethanol and its metabolites on the expression of AQPs. Materials & methods: CAPAN-1 cells were treated with ethanol (EtOH; 1-100 mM), chenodeoxycholate (CDC; 0.1-0.5 mM), glycochenodeoxycholate (GCDC; 0.1-0.5 mM) palmitoleic acid (POA; 10, 100 and 200 uM) and palmitoleic acid ethyl ester (POAEE; 10, 100 and 200 uM) for 6, 12, 24 and 48 hours and the expression of AQP isoforms (AQP1-12) was examined by real-time RT-PCR and immunocytochemistry. Results: All 12 AQPs were expressed in the CAPAN-1 cell line to a certain degree. AQP1, 3, 5, 6 and 11 were expressed at the highest levels while AQP2 and 4 were hardly detectable. In almost all treated group, the expression of AQPs decreased both at mRNA and protein levels dose- and time-dependently. Notably, a 72-hour incubation in culture media restored the expression of AQPs in the 6- and 12-hour CDC- and GCDC-treated groups and in the 24-hour EtOH-treated group. Conclusion: The role of AQP in the pathogenesis of AP needs further investigations. Our research is supported by Hungarian National Development Agency grants (TAMOP-4.2.2.A-11/1/KONV-2012-0035, -0052, -0073, TAMOP 4.2.4.A/2-11-1-2012-0001 ‘National Excellence Program) and the Hungarian Scientific Research Fund (OTKA NF105758, NF100677, K109756).
T-102. A human ex vivo system for the study of pancreatic injury and regeneration ndez Moro a, Sougat Misra a, Soledad Pouso a, Marita Carlos Ferna €hr c, Mikael Bjo €rnstedt a, Marco Wallenberg a, Rainer Heuchel b, Matthias Lo Del Chiaro c, Caroline Verbeke a a Division of Pathology, Department of Laboratory Medicine, Karolinska Institute, Stockholm, Sweden, Sweden b Department of Clinical Science, Intervention and Technology (CLINTEC), Center of Biosciences. Karolinska Institute, Stockholm, Sweden c Division of Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institute, Stockholm, Sweden
S79
Background: Animal experiments have shown that injury induces a regenerative response in pancreatic tissue, which is driven by the acinar compartment in the cerulein-induced pancreatitis model and by ductal epithelium in the ‘px model’. However, little is known about the response to injury in the human pancreas. Aims: To investigate regenerative responses in the human pancreas using an ex vivo culture system. Materials & methods: Fresh normal pancreatic tissue was sampled by knife-excision from surgical specimens (n¼10). The samples were sliced (350 mm thickness) and cultured for at least 4 days in organotypic culture under mildly hypoxic conditions (21% O2, no insert). Every 24 hours, two slices were processed for light microscopy, and sections were examined by two independent pathologists (CV, CFM) for signs of regeneration in the acinar and/or ductal compartment. Immunohistochemistry for epithelial (CK19, trypsin, Mist1), stromal (vimentin, aSMA), proliferative (Ki-67) and progenitor (PDX1, Hes1, SOX9) markers was performed. Results: From day 1, a single layer of epithelial cells was seen to grow out onto the surface of the tissue slice and appeared to originate from transected small branch ducts. During subsequent days, the cells extended progressively along the surface and acquired a duct-epithelial appearance (CK19+, vimentin). The acinar compartment showed acinar-ductal metaplasia, which was not connected with the cellular outgrowth on the tissue surface. Conclusion: The organotypic culture system shows evidence of injuryinduced duct-epithelial regeneration and acinar-ductal metaplasia.
T-103. Cigarette smoke extract inhibits fluid secretion and CFTR activity in guinea pig pancreatic ductal cells th a, Andrea Schnúr a, Jo zsef Mal Krisztina To eth a, Dezs Csupor b, c a eter Hegyi a Viktoria Venglovecz , Eleonora Gal , Zoltan Rakonczay, Jr. a, P a First Department of Medicine, University of Szeged, Szeged, Hungary, Hungary b Department of Pharmacognosy, University of Szeged, Szeged, Hungary, Hungary c Department of Pharmacology and Pharmacotherapy, University of Szeged, Szeged, Hungary, Hungary
Background: Smoking represents an independent risk factor for the development of chronic pancreatitis, however, the pathomechanism remains unknown. Secretion of fluid and bicarbonate plays a crucial role in maintaining the integrity of the gland. Aims: The aim of this study was to investigate the effects of cigarette smoke extract (CSE) on pancreatic ductal fluid secretion and on cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel activity. Materials & methods: Intra/interlobular pancreatic ducts were isolated from guinea pig pancreas. Basal and forskolin stimulated fluid secretion were measured by videomicroscopy, whereas, CFTR currents were detected by whole cell configuration of the patch clamp technique. CSE was prepared by smoking of 3 cigarettes into 40ml distilled water by a smoking machine and 10x (21mg/ml), 40x (5.25mg/ml) and 400x (0.5mg/ml) dilution of the extract were studied. Results: Administration of 5mM forskolin activated CFTR currents by 10-15-fold in magnitude. 15 min administration of 0.5, 5.25 and 21 mg/ml CSE inhibited the currents by 44%, 64.6% and 79.4%, respectively (n¼2-4). Concerning the fluid secretion, the basal volume of isolated intact pancreatic ducts in bicarbonate-free solution was considered to be 1.0. Administration of 25mM bicarbonate increased the relative luminal volume up to 1.57±0.02 (n¼7). Administration of 5 mM forskolin further increased the luminal volume to 1.87±0.1 (n¼16). Simultaneous administration of 21mg/ml CSE decreased fluid secretion by 24% (1.42±0.06; n¼12).
Abstracts / Pancreatology 14 (2014) S1eS129
Background: There are no extensive data on the comparative evaluation of 25-hydroxyvitamin D (VIT D) serum levels in patients with chronic benign and malignant pancreatic diseases. Aims: To evaluate serum VIT D levels in patients with chronic pancreatitis (CP), pancreatic adenocarcinoma (PA) and in healthy subjects (HS). Patients & methods: Sixty-nine consecutive subjects were studied: 20 CP patients (12 M, 8 F; mean age 56.0 yrs, range 25-74; 14 were alcohol drinkers and 12, smokers; 12 had calcifications and 5 diabetes mellitus), 20 PA (11 M, 9 F; age 63.5 yrs, range 47-80; 5 patients were alcohol drinkers, 3 were smokers; 11 had diabetes); 29 HS (13 M, 16 F; mean age 59.5 yrs, range 44-75). Serum VIT D levels were measured using chemiluminescence assay . VIT D levels were normally distributed and ANOVA test was applied. Fecal pancreatic elastase-1 (FE) was also measured in patients. Results: The 3 groups were comparable for sex (P¼0.554) and age (P¼0.107); 14 CP patients and 1 PA had exocrine pancreatic insufficiency (P<0.001). Serum VIT D was significantly different among the 3 groups of patients studied (mean±SD; CP: 10.9±8.1 ng/mL, PA: 12.5±5.6 ng/mL, HS: 23.6±8.3 ng/mL; P<0.001). CP and PA patients had similar VIT D serum levels (P¼0.258) and they were significantly lower than in HS (P<0.001). At univariate analysis only the type of the disease (i.e. CP or PA) was significantly associated with low VIT D levels (P¼0.036). Conclusion: Low VIT D levels are associated with chronic diseases of the pancreas, but the mechanism should be better elucidated.
Aims: This study aims to distinguish nutritional status of TP-IAT patients pre- and post-surgery. Patients & methods: Nutrition parameters (BMI, vitamin D3, vitamin A, and vitamin E levels) were obtained from an IRB-approved prospective database collected for TP-IAT patients at a single institution. Two-tailed paired T-test and two-tailed student T-test for unequal variances were used for data analysis. Results: Weight loss was observed from pre-op to 6 months post-op (n¼111) from mean BMI (26.1kg/m2+/-6.6 to 23.0kg/m2+/-4.9, p<0.0001). There was little change in weight from 6 months to 1 year post-op. BMI at 2 and 3 years post-op were also unchanged. Average vitamin D3 levels dropped significantly pre-op to 6 months post-op (27.8ng/ml+/-10.5 to 24.4ng/ml+/-10.2, p¼0.02). Levels also dropped from 6 months post-op to 1 year post-op (24.4ng/ml+/-10.2 to 22.5ng/ml+/-11.8) indicating moderate deficiency after TP-IAT. Despite having fewer data at 2 and 3 years post-op, mean vitamin D3 levels indicated prolonged vitamin D3 deficiency (20.3ng/ml+/-11.8, 21.6ng/ml+/-15.8). Mean levels of a subpopulation (n¼55) for vitamin A and vitamin E were normal (0.34mg/L+/-0.15 and 7.4mg/L+/-3.1) at time points ranging from 6 months post-op to 3 years post-op. However, vitamin A and vitamin E deficiencies were observed in 45% and 29% of subjects respectively. Conclusion: Nutritional compromise is a notable problem in patients after TP-IAT. Although nutritional risk may improve over time, these patients require long-term follow-up.
T-098. Clinical significance of sarcopenia as a predictor of prognosis in patients who underwent pancreaticoduodenectomy. Fumiaki Watanabe, Hiroshi Noda, Toshiki Rikiyama Jichi Medical University Saitama Medical Center, Surgery, Japan Background: Sarcopenia was identified recently as a poor prognostic factor in patients with cancer. The present study investigated the influence of sarcopenia on the perioperative clinical outcomes and long-term survival of the patients who underwent pancreaticoduodenectomy(PD). Aims: Patients were assigned to one of two groups according to the presence or absence of sarcopenia. Clinicopathological, surgical outcome and long-term survival data were analysed. Patients & methods: Data were collected retrospectively for 123 consecutive patients who underwent PD for pancreatic head cancer, carcinoma of vater, common bile duct cancer, and IPMN with a curative intent between April 2008 and December 2012. Patients were divided into two groups according to the presence or absence of sarcopenia, assessed by computed tomographic measurement of muscle mass at the level of the third lumbar vertebra (N. Harimoto, et al. British Journal of Surgery 2013; 100: 1523e30). Clinicopathologica characteristics, postoperative outcomes and long-term survival data were compared between two groups. Results: Sarcopenia was present in 60 (48.8 per cent) of 123 patients, and was significantly correlated with female sex. In patients with and without sarcopenia, the rate of morbidities was 30.0 and 36.5 per cent, the duration postoperative hospital stay was 29.0 and 29.3 days, and the 5-year overall survival rate was 54.3 and 76.7 per cent, respectively (p¼0.067). Conclusion: It might be possible that sarcopenia was a predictive factor for poor prognosis of patients who underwent PD.
T-099. Auto-islet cell transplantation: What is happening to weight and fatsoluble vitamin status? Maria Nestleroad, Kelley Martin, Elizabeth Shufford, Stefanie Owczarski, David Adams, Katherine Morgan Medical University of South Carolina, Department of Gastrointestinal and Laparoscopic Surgery, United States Background: Limited data links chronic pancreatitis to nutritional compromise after Total pancreatectomy and islet auto-transplantation (TPIAT).
T-100. A common SLC26A6 haplotype is not associated with chronic pancreatitis zs a, Eszter Hegyi a, Bala zs Csaba N n Hritz c, Judit Anita Bala emeth b, Istva c c d d € Gervain , Ferenc Izbeki , Attila Szepes , Gyorgy Gyimesi , Dezs g, Zsolt Vincze e, Judit Bajor f, Adrienn Csiszko Kelemen e, Aron s Bod h, Andrea Pa rniczky i, La sztity Nata lia i, Szentkereszty g, Barnaba bor Veres j, Richa rd Szmola k, Akos Pap k , Judit Czelecz l, Csilla Andorka j, Ga nos Nova k n, Gyula Farkas o, La szlo s nos Sümegi m, Ja Czako a, Tama Ja a a a th b, P eter Takacs , Zoltan Rakonczay , Jozsef Maleth , Miklos Sahin-To Hegyi a a
First Department of Medicine, University of Szeged, Szeged, Hungary Department of Molecular and Cell Biology, Boston University, Boston, United States c r Megyei Szent Gyo € rgy Hospital, Sze kesfehe rva r, Hungary Feje d t, Hungary Bacs-Kiskun County Municipality Hospital, Kecskeme e cs, Hungary Department of Surgery, University of Pe f cs, Hungary Department of Internal Medicine, University of Pe g Department of Surgery, University of Debrecen, Hungary h n Hospital, Szentes, Hungary Dr. Bugyi Istva i Heim P al Children?s Hospital, Budapest, Hungary j Semmelweis University,Paediatric Department, Budapest, Hungary k National Institute of Oncology, Budapest, Hungary, Hungary l Bethasda Children?s Hospital, Budapest, Hungary m B-A-Z County Hopspital, Miskolc, Hungary n ndy Ka lma n County Hospital, Gyula, Hungary 9Pa o Department of Surgery, University of Szeged, Hungary b
Background: Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) mutations are established risk factors for chronic pancreatitis (CP). CFTR variants increase disease risk by causing impairment of pancreatic ductal bicarbonate secretion. However, the role of genetic variations in the bicarbonate secreting SLC26A6 anion transporter has remained largely unexplored so far. Aims: Our aim was to investigate the role of the SLC26A6 gene in CP. Patients & methods: 96 subjects with CP (cases) and 99 subjects with no pancreatic disease (controls) were recruited from the Hungarian National Pancreas Registry. In a discovery cohort of 30 idiopathic CP cases the entire SLC26A6 coding sequence, including 21 exons and the exonintron boundaries were amplified and sequenced. Further genotyping of p.V206M and p.P397P mutations in CP and controls was carried out by RFLP.
Abstracts / Pancreatology 14 (2014) S1eS129
Results: Sequencing analysis of the discovery cohort revealed four common mutations: intronic mutations c.23+71_23+103del, c.183-4C>A and c.1134+32C>A; and exonic missense mutation p.V206M. . These four mutations were found in linkage disequilibrium indicating a conserved haplotype. We found this haplotype in 18 heterozygous and 2 homozygous cases, and in 24 heterozygous and 2 homozygous controls (allele frequency 11.4% and 14.1% respectively). A synonymous mutation p.P397P was also detected in a single case. Conclusion: We found a novel, common haplotype in the SLC26A6 gene, which did not show association with CP. Supported by TAMOP and OTKA.
T-101. Expression of aquaporins in pancreatic ductal cells n Rakonczay, Jr. b, Viktoria Venglovecz a, Lajos V. Kem eny b, Zolta szlo s c, P Puska eter Hegyi b Agnes Zvara c, La a University of Szeged, Department of Pharmacology and Pharmacotherapy, Hungary b University of Szeged, First Department of Medicine, Hungary c Biological Research Centre, Laboratory of Functional Genomics, Hungary
Background: Acute pancreatitis (AP) is a multicellular disease in which pancreatic ductal cells play an important role. Toxic agents inducing AP inhibit pancreatic ductal bicarbonate secretion, however, no information is available concerning their effects on the regulation of aquaporins (AQPs). Aims: The aim of this study was to investigate the effects of bile acids, ethanol and its metabolites on the expression of AQPs. Materials & methods: CAPAN-1 cells were treated with ethanol (EtOH; 1-100 mM), chenodeoxycholate (CDC; 0.1-0.5 mM), glycochenodeoxycholate (GCDC; 0.1-0.5 mM) palmitoleic acid (POA; 10, 100 and 200 uM) and palmitoleic acid ethyl ester (POAEE; 10, 100 and 200 uM) for 6, 12, 24 and 48 hours and the expression of AQP isoforms (AQP1-12) was examined by real-time RT-PCR and immunocytochemistry. Results: All 12 AQPs were expressed in the CAPAN-1 cell line to a certain degree. AQP1, 3, 5, 6 and 11 were expressed at the highest levels while AQP2 and 4 were hardly detectable. In almost all treated group, the expression of AQPs decreased both at mRNA and protein levels dose- and time-dependently. Notably, a 72-hour incubation in culture media restored the expression of AQPs in the 6- and 12-hour CDC- and GCDC-treated groups and in the 24-hour EtOH-treated group. Conclusion: The role of AQP in the pathogenesis of AP needs further investigations. Our research is supported by Hungarian National Development Agency grants (TAMOP-4.2.2.A-11/1/KONV-2012-0035, -0052, -0073, TAMOP 4.2.4.A/2-11-1-2012-0001 ‘National Excellence Program) and the Hungarian Scientific Research Fund (OTKA NF105758, NF100677, K109756).
T-102. A human ex vivo system for the study of pancreatic injury and regeneration ndez Moro a, Sougat Misra a, Soledad Pouso a, Marita Carlos Ferna €hr c, Mikael Bjo €rnstedt a, Marco Wallenberg a, Rainer Heuchel b, Matthias Lo Del Chiaro c, Caroline Verbeke a a Division of Pathology, Department of Laboratory Medicine, Karolinska Institute, Stockholm, Sweden, Sweden b Department of Clinical Science, Intervention and Technology (CLINTEC), Center of Biosciences. Karolinska Institute, Stockholm, Sweden c Division of Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institute, Stockholm, Sweden
S79
Background: Animal experiments have shown that injury induces a regenerative response in pancreatic tissue, which is driven by the acinar compartment in the cerulein-induced pancreatitis model and by ductal epithelium in the ‘px model’. However, little is known about the response to injury in the human pancreas. Aims: To investigate regenerative responses in the human pancreas using an ex vivo culture system. Materials & methods: Fresh normal pancreatic tissue was sampled by knife-excision from surgical specimens (n¼10). The samples were sliced (350 mm thickness) and cultured for at least 4 days in organotypic culture under mildly hypoxic conditions (21% O2, no insert). Every 24 hours, two slices were processed for light microscopy, and sections were examined by two independent pathologists (CV, CFM) for signs of regeneration in the acinar and/or ductal compartment. Immunohistochemistry for epithelial (CK19, trypsin, Mist1), stromal (vimentin, aSMA), proliferative (Ki-67) and progenitor (PDX1, Hes1, SOX9) markers was performed. Results: From day 1, a single layer of epithelial cells was seen to grow out onto the surface of the tissue slice and appeared to originate from transected small branch ducts. During subsequent days, the cells extended progressively along the surface and acquired a duct-epithelial appearance (CK19+, vimentin). The acinar compartment showed acinar-ductal metaplasia, which was not connected with the cellular outgrowth on the tissue surface. Conclusion: The organotypic culture system shows evidence of injuryinduced duct-epithelial regeneration and acinar-ductal metaplasia.
T-103. Cigarette smoke extract inhibits fluid secretion and CFTR activity in guinea pig pancreatic ductal cells th a, Andrea Schnúr a, Jo zsef Mal Krisztina To eth a, Dezs Csupor b, c a eter Hegyi a Viktoria Venglovecz , Eleonora Gal , Zoltan Rakonczay, Jr. a, P a First Department of Medicine, University of Szeged, Szeged, Hungary, Hungary b Department of Pharmacognosy, University of Szeged, Szeged, Hungary, Hungary c Department of Pharmacology and Pharmacotherapy, University of Szeged, Szeged, Hungary, Hungary
Background: Smoking represents an independent risk factor for the development of chronic pancreatitis, however, the pathomechanism remains unknown. Secretion of fluid and bicarbonate plays a crucial role in maintaining the integrity of the gland. Aims: The aim of this study was to investigate the effects of cigarette smoke extract (CSE) on pancreatic ductal fluid secretion and on cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel activity. Materials & methods: Intra/interlobular pancreatic ducts were isolated from guinea pig pancreas. Basal and forskolin stimulated fluid secretion were measured by videomicroscopy, whereas, CFTR currents were detected by whole cell configuration of the patch clamp technique. CSE was prepared by smoking of 3 cigarettes into 40ml distilled water by a smoking machine and 10x (21mg/ml), 40x (5.25mg/ml) and 400x (0.5mg/ml) dilution of the extract were studied. Results: Administration of 5mM forskolin activated CFTR currents by 10-15-fold in magnitude. 15 min administration of 0.5, 5.25 and 21 mg/ml CSE inhibited the currents by 44%, 64.6% and 79.4%, respectively (n¼2-4). Concerning the fluid secretion, the basal volume of isolated intact pancreatic ducts in bicarbonate-free solution was considered to be 1.0. Administration of 25mM bicarbonate increased the relative luminal volume up to 1.57±0.02 (n¼7). Administration of 5 mM forskolin further increased the luminal volume to 1.87±0.1 (n¼16). Simultaneous administration of 21mg/ml CSE decreased fluid secretion by 24% (1.42±0.06; n¼12).